Rat liver DT-diaphorase: regulation of functional mRNA levels by 3-methylcholanthrene, trans-stilbene oxide, and phenobarbital

Arch Biochem Biophys. 1984 Jul;232(1):408-13. doi: 10.1016/0003-9861(84)90556-3.


Total liver poly(A+)-RNA isolated from untreated, and 3-methylcholanthrene-, trans-stilbene oxide-, and phenobarbital-treated rats has been translated in the rabbit reticulocyte lysate system in order to determine the effect of these xenobiotics on the level of translationally active DT-diaphorase mRNA. The in vitro translation systems were subjected to immunoprecipitation with rabbit IgG raised against purified DT-diaphorase and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identity of the radiolabeled, immunoprecipitated product as DT-diaphorase was confirmed by limited peptide mapping using Staphylococcus aureus V-8 protease. These quantitation results demonstrate that 3-methylcholanthrene leads to an eight-fold elevation in functional DT-diaphorase mRNA at 8 h after a single administration of 3-methylcholanthrene; whereas, trans-stilbene oxide and phenobarbital produced only a modest elevation, two- to three-fold, in the functional DT-diaphorase mRNA level. These data indicate that the increase in the level of DT-diaphorase after 3-methylcholanthrene administration noted previously [B. Höjeberg, K. Blomberg, S. Stenberg, and C. Lind (1981) Arch. Biochem. Biophys. 207, 205-216] can be totally accounted for by an elevation in the mRNA level specific for this protein.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell-Free System
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Induction / drug effects
  • Immunochemistry
  • Liver / enzymology*
  • Methylcholanthrene / pharmacology*
  • NAD(P)H Dehydrogenase (Quinone)
  • NADH, NADPH Oxidoreductases / biosynthesis*
  • Phenobarbital / pharmacology*
  • Quinone Reductases / biosynthesis*
  • Quinone Reductases / genetics
  • RNA, Messenger / biosynthesis*
  • Rabbits
  • Rats
  • Stilbenes / pharmacology*


  • RNA, Messenger
  • Stilbenes
  • Methylcholanthrene
  • NADH, NADPH Oxidoreductases
  • NAD(P)H Dehydrogenase (Quinone)
  • Quinone Reductases
  • stilbene oxide
  • Phenobarbital